Abstract:

Abstract:Objective To construct the human BRCA1 promotor luciferase report gene vector and detect its activity in cells. Methods The BRCA1 promoter from human normal cervix tissues                 
  was amplified by PCR, and was inserted into the luciferase report gene pGL3-basic vector. The amplified DNA sequence was confirmed by sequencing and then the constructed vector was transfected into  HCT116 cells to detect its activity by Premaga Dual-luciferase report gene detection system. Results The recombinant plasmid was tested by gel electrophoresis and sequencing analysis, it was proved that the plasmid included pGL3-basic DNA sequence and PRL regulating sequence.The sequencing results indicated that the amplified sequence was correct, in p53 minus HCT116 cells the number of BRCA1 promoter was increased (P<0.05), and the luciferase activity detection result demonstrated that the constructed vector had the promotor activity.Conclusion The human BRCA1 promotor luciferase report gene vector has been constructed successfully, and it will become essential material for further study on the function of BRCA1 regulation.

Key words: BRCA1, promotor, luciferase, report gene